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Literature summary for 1.12.1.2 extracted from

  • Schneider, K.; Schlegel, H.G.; Jochim, K.
    Effect of nickel on activity and subunit composition of purified hydrogenase from Nocardia opaca 1 b (1984), Eur. J. Biochem., 138, 553-541.
No PubMed abstract available
Search for more literature for 1.12.1.2

General Stability

General Stability Organism
Ni2+ stabilizes Rhodococcus opacus

Inhibitors

Inhibitors Comment Organism Structure
Ni2+ 0.5-1 mM, strong inhibition of artificial electron acceptor reduction Rhodococcus opacus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.05
-
 methyl viologen at pH 9.4 Rhodococcus opacus
0.15
-
 NAD+
-
 Rhodococcus opacus
0.38
-
 FAD
-
 Rhodococcus opacus
0.44
-
 methylene blue
-
 Rhodococcus opacus
0.9
-
 methyl viologen at pH 8.0 Rhodococcus opacus
4
-
 ferricyanide
-
 Rhodococcus opacus

Localization

Localization Comment Organism GeneOntology No. Textmining
soluble
-
 Rhodococcus opacus
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ NAD+ reduction with H2 is completely dependent on the presence of divalent metal ions Ni2+, Co2+, Mg2+ or Mn2+ or of high salt concentrations between 500-1500 mM Rhodococcus opacus
Mg2+ NAD+ reduction with H2 is completely dependent on the presence of divalent metal ions Ni2+, Co2+, Mg2+ or Mn2+ or of high salt concentrations between 500-1500 mM Rhodococcus opacus
Mn2+ NAD+ reduction with H2 is completely dependent on the presence of divalent metal ions Ni2+, Co2+, Mg2+ or Mn2+, or of high salt concentrations of 500-1500 mM Rhodococcus opacus
additional information stimulation of activity by salt is greater the less chaotrophic the anion Rhodococcus opacus
Ni2+ NAD+ reduction with H2 is completely dependent on the presence of divalent metal ions Ni2+, Co2+, Mg2+ or Mn2+ or of high salt concentrations between 500-1500 mM Rhodococcus opacus
Ni2+ highest specific activity with NiCl2, optimal concentration: 1 mM Rhodococcus opacus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
27000
-
 1 * 64000 + 1 * 56000 + 1 * 31000 + 1 * 27000, native hydrogenase dissociates into two subunit dimers of 64000 Da and 1 * 31000 Da, and of 56000 Da and 27000 Da respectively, SDS-PAGE Rhodococcus opacus
31000
-
 1 * 64000 + 1 * 56000 + 1 * 31000 + 1 * 27000, native hydrogenase dissociates into two subunit dimers of 64000 Da and 1 * 31000 Da, and of 56000 Da and 27000 Da respectively, SDS-PAGE Rhodococcus opacus
56000
-
 1 * 64000 + 1 * 56000 + 1 * 31000 + 1 * 27000, native hydrogenase dissociates into two subunit dimers of 64000 Da and 1 * 31000 Da, and of 56000 Da and 27000 Da respectively, SDS-PAGE Rhodococcus opacus
64000
-
 1 * 64000 + 1 * 56000 + 1 * 31000 + 1 * 27000, native hydrogenase dissociates into two subunit dimers of 64000 Da and 1 * 31000 Da, and of 56000 Da and 27000 Da respectively, SDS-PAGE Rhodococcus opacus
178000
-
 sucrose density gradient centrifugation Rhodococcus opacus
200000
-
 gel filtration Rhodococcus opacus

Organism

Organism UniProt Comment Textmining
Rhodococcus opacus
-
-
-
Rhodococcus opacus 1B
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Rhodococcus opacus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
45
-
-
 Rhodococcus opacus

Storage Stability

Storage Stability Organism
4°C, 500 mM potassium phosphate, pH 7.0, 3 d, 10% loss of activity Rhodococcus opacus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
FAD + H2 99% activity compared to NAD+ Rhodococcus opacus FADH2
-
 r
H2 + NAD+ electron acceptor FAD Rhodococcus opacus H+ + NADH
-
 r
H2 + NAD+ artificial electron acceptor benzyl viologen Rhodococcus opacus H+ + NADH
-
 r
H2 + NAD+ artificial electron acceptor methyl viologen Rhodococcus opacus H+ + NADH
-
 r
H2 + NAD+ electron acceptor NAD+ Rhodococcus opacus H+ + NADH
-
 r
H2 + NAD+ artificial electron acceptor ferricyanide Rhodococcus opacus H+ + NADH
-
 r
H2 + NAD+ artificial electron acceptor methylene blue Rhodococcus opacus H+ + NADH
-
 r
H2 + NAD+ electron acceptor FAD Rhodococcus opacus 1B H+ + NADH
-
 r
H2 + NAD+ artificial electron acceptor benzyl viologen Rhodococcus opacus 1B H+ + NADH
-
 r
H2 + NAD+ artificial electron acceptor methyl viologen Rhodococcus opacus 1B H+ + NADH
-
 r
H2 + NAD+ electron acceptor NAD+ Rhodococcus opacus 1B H+ + NADH
-
 r
H2 + oxidized benzyl viologen 50fold higher specific activity compared to NAD+ reduction Rhodococcus opacus H+ + reduced benzyl viologen
-
 r
H2 + oxidized methyl viologen
-
 Rhodococcus opacus H+ + reduced methyl viologen
-
 ?
H2 + oxidized methyl viologen
-
 Rhodococcus opacus 1B H+ + reduced methyl viologen
-
 ?
H2 + oxidized methylene blue
-
 Rhodococcus opacus H+ + reduced methylene blue
-
 r
oxidized methylene blue + H2 1058% activity compared to NAD+ Rhodococcus opacus reduced methylene blue + H+
-
 r

Subunits

Subunits Comment Organism
tetramer 1 * 64000 + 1 * 56000 + 1 * 31000 + 1 * 27000, native hydrogenase dissociates into two subunit dimers of 64000 Da and 1 * 31000 Da, and of 56000 Da and 27000 Da respectively, SDS-PAGE Rhodococcus opacus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 electron donor NADH Rhodococcus opacus
6.1
-
 electron acceptor NAD+, no addition of Ni2+, Tris/MES buffer Rhodococcus opacus
7.8 8 electron acceptor NAD+, in triethanolamine/HCl buffer containing NiCl2 Rhodococcus opacus

pH Range

pH Minimum pH Maximum Comment Organism
additional information
-
-
 Rhodococcus opacus

pH Stability

pH Stability pH Stability Maximum Comment Organism
6
-
 1 day: 9% loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 56% loss of activity in the absence of NiCl2 and MgCl2, 3 days: 44% loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 80% loss of activity in the absence of NiCl2 and MgCl2 Rhodococcus opacus
7
-
 1 day: no loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 62% loss of activity in the absence of NiCl2 and MgCl2, 3 days: 21% loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 87% loss of activity in the absence of NiCl2 and MgCl2 Rhodococcus opacus
8
-
 1 day: no loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 86% loss of activity in the absence of NiCl2 and MgCl2, 3 days: 27% loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, complete loss of activity in the absence of NiCl2 and MgCl2 Rhodococcus opacus